Identification of the DDAH2 Protein in Pig Reproductive Tract Mucus: A Putative Oestrus Detection Marker

Precisely detecting oestrus is important for artificial insemination. The aims of this study were to identify oestrus‐specific sow mucus proteins to determine the optimal time for artificial insemination. The proestrous‐ and oestrous‐stage mucus proteins were purified and analysed with proteomic tools such as two‐dimensional gel electrophoresis and matrix‐assisted laser desorption/ionization–time‐of‐flight analyses. Among the differentially expressed proteins, the dimethylarginine dimethylaminohydrolase 2 (DDAH2) protein showed a 3.6‐fold increase during the proestrous stage compared to that during the oestrous stage. A western immunoblot study revealed that two of three sow mucus samples clearly showed negative anti‐DDAH2 antibody activity during the oestrous stage. This study demonstrated that the pig DDAH2 mucus protein exists during the proestrous stage, but not during the oestrous stage, suggesting that mucus DDAH2 could be useful as an oestrus detection marker.     

The effect of reduced dose and number of treatments of FSH on superovulatory response in CIDR-treated Korean native cows

The objective of this study was to investigate the effect of dosage and number of days of follicle stimulating hormone (FSH) treatment on superovulatory response in controlled internal drug release (CIDR)-treated Korean native cows. Forty cows underwent two superovulatory treatments with a crossover design. Cows, at random stages of the estrous cycle, received a CIDR together with injections of 1 mg estradiol benzoate and 50 mg progesterone, and gonadotropin treatment began 4 days later. The cows were divided into 2 groups based on the dosage and number of days of treatment with porcine FSH; a total of 28 mg FSH was given in twice daily intramascular injections in decreasing doses over 4 days (5, 5, 4, 4, 3, 3, 2 and 2 mg; T1 group, n=20) or a total of 24 mg FSH was given in twice daily decreasing doses over 3 days (5, 5, 4, 4, 3 and 3 mg; T2 group, n=20). This was followed by the alternate treatment in the subsequent superovulation. The cows were treated identically in all other respects. PGF(2alpha) (25 mg and 15 mg) was given with the 5th and 6th injections of FSH, CIDR were withdrawn at the 6th FSH injection and the cows received 200 microg GnRH 36 h after CIDR withdrawal. The cows were artificially inseminated twice, at 48 and 60 h after CIDR withdrawal, using commercial semen from four Korean native bulls, and embryos were recovered 6 or 7 days after the 2nd insemination. The numbers of corpora lutea (CL; 7.9+/-1.0 vs. 8.3+/-1.1) and large follicles (1.2+/-0.2 vs. 1.3+/-0.3) present at the time embryo recovery, as detected by ultrasonography, did not differ between the T1 and T2 groups (P>0.05). Similarly, the numbers of total ova/embryos (6.2+/-0.9 vs. 6.4+/-1.1), transferable embryos (3.4+/-0.8 vs. 3.2+/-0.7), degenerate embryos (0.8+/-0.2 vs. 1.0+/-0.3) and unfertilized ova (2.1+/-0.5 vs. 2.2+/-0.5) did not differ between the groups (P>0.05). These data indicate that a reduced dose (24 vs. 28 mg) and number of treatments (6 vs. 8) of FSH for superovulation of CIDR-treated Korean native cows does not affect the embryo yield.     

NMR structure of Mistic, a membrane-integrating protein for membrane protein expression

Although structure determination of soluble proteins has become routine, our understanding of membrane proteins has been limited by experimental bottlenecks in obtaining both sufficient yields of protein and ordered crystals. Mistic is an unusual Bacillus subtilis integral membrane protein that folds autonomously into the membrane, bypassing the cellular translocon machinery. Using paramagnetic probes, we determined by nuclear magnetic resonance (NMR) spectroscopy that the protein forms a helical bundle with a surprisingly polar lipid-facing surface. Additional experiments suggest that Mistic can be used for high-level production of other membrane proteins in their native conformations, including many eukaryotic proteins that have previously been intractable to bacterial expression.     

An Alternate Method for Fourier Transform Infrared (FTIR) Spectroscopic Determination of Soil Nitrate Using Derivative Analysis and Sample Treatments

This study aimed at examining effective sample treatments and spectral processing for an alternate method of soil nitrate determination using the attenuated total reflectance (ATR) of Fourier transform infrared (FTIR) spectroscopy. Prior to FTIR measurements, soil samples were prepared as paste to enhance adhesion between the ATR crystal and sample. The similar nitrate peak heights of soil pastes and their supernatants indicated that the nitrate in the liquid portion of the soil paste mainly responded to the FTIR signal. Using a 0.01-M CaSO₄ solution for the soil paste, which has no interference bands in the characteristic spectra of the analyte, increased the concentration of the nitrates to be measured. Second-order derivatives were used in the prediction model to minimize the interference effects and enhance the performance. The second-order derivative spectra contained a unique nitrate peak in a range of 1,400–1,200 cm^?1 without interference of carbonate. A partial least square regression model using second-order derivative spectra performed well (R ²?=?0.995, root mean square error (RMSE)?=?23.5, ratio of prediction to deviation (RPD)?=?13.8) on laboratory samples. Prediction results were also good for a test set of agricultural field soils with a CaCO₃ concentration of 6% to 8% (R ²?=?0.97, RMSE?=?18.6, RPD?=?3.5). Application of the prediction model based on soil paste samples to nitrate stock solution resulted in an increased RMSE (62.3); however, validation measures were still satisfactory (R ²?=?0.99, RPD?=?3.0).     

Genetic and QTL analysis of pericarp thickness and ear architecture traits of Korean waxy corn germplasm

Pericarp thickness and ear traits are important selection criteria for breeding fresh market waxy corn. This research was conducted to better understand genetic control of these traits in popular South Korean germplasm now grown in Illinois. Pericarp thickness on five kernel regions, and ten inflorescence architecture traits were measured on ears from 264 F_2:3 families from a cross between Korean inbreds BH20 and BH30. All five pericarp thickness traits showed high heritabilities and were highly correlated. Multivariate principal components analysis (PCA) revealed that just one principal component (PC) explained most of the total phenotypic variation. A number of univariate quantitative trait loci (QTL) were detected and most were associated with more than one kernel pericarp region. Four out of seven PC-QTL were located in chromosome positions where three or more pericarp thickness univariate QTL were detected. Conversely, three PC-QTL were found in regions with just a single or two univariate QTL, indicating that these QTL regions may be more important for overall pericarp thickness than suggested by univariate analysis. The PCA, QTL, and PC-QTL results indicate that pericarp thickness on different kernel regions may be controlled by common genes with pleiotropic effects. Additive effects of QTL for thinner pericarp thickness came from both BH20 and BH30. For ear architecture traits, heritability varied from 0.38 to 0.72, and several traits were correlated. The PCA reduced these traits into three independent PCs, and all substantial component traits for these PCs were also significantly correlated. A number of univariate QTL were clustered closely, and some PC-QTL were detected in these regions. Some PC-QTL were found in chromosome regions where univariate QTL were not detected, again suggesting that these regions may have larger overall effects on ear architecture than suggested by univariate analyses. Collectively, these QTL may be useful for marker assisted introgression into germplasm more adapted to the U.S.     

Egg production, faecal pH and microbial population, small intestine morphology, and plasma and yolk cholesterol in laying hens given liquid metabolites produced by Lactobacillus plantarum strains

1. Various dosages of metabolite combinations of the Lactobacillus plantarum RI11, RG14 and RG11 strains (COM456) were used to study the egg production, faecal microflora population, faecal pH, small intestine morphology, and plasma and egg yolk cholesterol in laying hens. 2. A total of 500 Lohmann Brown hens were raised from 19 weeks to 31 weeks of age. The birds were randomly divided into 5 groups and fed on various treatment diets: (i) basal diet without supplementation of metabolites (control); (ii) basal diet supplemented with 0·3% COM456 metabolites; (iii) basal diet supplemented with 0·6% COM456 metabolites; (iv) basal diet supplemented with 0·9% COM456 metabolites; and (v) basal diet supplemented with 1·2% COM456 metabolites. 3. The inclusion of 0·6% liquid metabolite combinations, produced from three L. plantarum strains, demonstrated the best effect in improving the hens' egg production, faecal lactic acid bacteria population, and small intestine villus height, and reducing faecal pH and Enterobacteriaceae population, and plasma and yolk cholesterol concentrations. 4. The metabolites from locally isolated L. plantarum are a possible alternative feed additive in poultry production.     

Phylogenetic analysis of benign Theileria species based on major piroplasm surface protein (MPSP) genes from ticks of grazing cattle in Korea

Complete major piroplasm surface protein (MPSP) gene sequences of benign Theileria parasites were isolated from ticks of grazing cattle in Korea. A total of 556 tick samples were collected in five provinces: Chungbuk, Jeonbuk, Jeonnam, Gyeongbuk, and Jeju during 2010-2011. Fifteen samples from Chungbuk and Jeonnam were positive for the Theileria MPSP gene by PCR amplification using a specific primer set. A phylogenetic tree was constructed with the amplified gene sequences and 26 additional sequences published in GenBank. The benign Theileria parasites were classified into eight types, those isolated from Korean cattle ticks belonged to Types 1 (Ikeda), 2 (Chitose), 4, and 8. Types 2 and 4 were the most common types, with the rate of 40%, followed by Types 1 and 8 (with the rate of 13% and 7%, respectively). Nucleotide sequence identities of 23 theilerial MPSP sequences (15 MPSP gene sequences amplified and 8 sequences published) ranged from 67.3 to 99.8%. Multiple alignments of the deduced amino acid sequences also showed that each type was characterized by specific amino acids: 7 for Type 1, 9 for Type 2, 4 for Type 4, and 3 for Type 8.     

Modification of ascorbic acid using transglycosylation activity of Bacillus stearothermophilus maltogenic amylase to enhance its oxidative stability

Ascorbic acid (1), a natural antioxidant, was modified by employing transglycosylation activity of Bacillus stearothermophilus maltogenic amylase with maltotriose and acarbose as donor molecules to enhance its oxidative stability. The transglycosylation reaction with maltotriose as donor created mono- and di-glycosyl transfer products with an alpha-(1,6)-glycosidic linkage. In addition, two acarviosine-glucosyl transfer products were generated when transglycosylation was performed with acarbose as a donor. All transfer products were observed by TLC and HPLC, and purified by Q-sepharose anion exchange and Biogel P-2 gel permeation chromatographies. LC/MS and (13)C NMR analyses revealed that the structures of the transfer products were 6-O-alpha-D-glucosyl- (2) and 6-O-alpha-D-maltosyl-ascorbic acids (3) in the reaction of maltotriose, and 6-O-alpha-acarviosine-D-glucosyl- (4) and 2-O-alpha-acarviosine-D-glucosyl ascorbic acids (5) in the reaction of acarbose. The stability of the transglycosylated ascorbic acid derivatives was greatly enhanced against oxidation by Cu(2+) ion and ascorbate oxidase. Among them, compound 3 proved to be the most stable against in vitro oxidation. The antioxidant effects of glycosyl-derivatives of ascorbic acid on the lipid oxidation in cooked chicken breast meat patties indicated that they had antioxidant activities similar to that of ascorbic acid. It is suggested that the transglycosylated ascorbic acids can possibly be applied as effective antioxidants with improved stability in food, cosmetic, and other applications.     

Vortex core-driven magnetization dynamics

Time-resolved x-ray imaging shows that the magnetization dynamics of a micron-sized pattern containing a ferromagnetic vortex is determined by its handedness, or chirality. The out-of-plane magnetization in the nanometer-scale vortex core induces a three-dimensional handedness in the planar magnetic structure, leading to a precessional motion of the core parallel to a subnanosecond field pulse. The core velocity was an order of magnitude higher than expected from the static susceptibility. These results demonstrate that handedness, already well known to be important in biological systems, plays an important role in the dynamics of microscopic magnets.